微小RNA-499-5p靶向CYLD对肿瘤细胞增殖和侵袭的影响  

The effect of microRNA-499-5p targeting CYLD on tumor cell proliferation and invasion

作　　者：高玉梅[1] 冯兴梅[1] 侯妍妍 李飞[2] Gao Yumei;Feng Xingmei;Hou Yanyan;Li Fei(Department of Gynecologic Oncology,the First People’s Hospital of Shangqiu,Shangqiu 476100,China;Department of Reproductive Medicine,the First People’s Hospital of Shangqiu,Shangqiu 476100,China)

机构地区：[1]商丘市第一人民医院(徐州医科大学商丘临床学院)妇瘤科,商丘476100 [2]商丘市第一人民医院(徐州医科大学商丘临床学院)生殖医学科,商丘476100

出　　处：《中华实验外科杂志》2023年第12期2566-2569,共4页Chinese Journal of Experimental Surgery

基　　金：河南省医学科技攻关联合共建项目(LHGJ20200933)。

摘　　要：目的探讨微小RNA(miRNA,miR)-499-5p靶向CYLD对肿瘤细胞增殖和侵袭的影响。方法选取2020年5月至2022年5月我院收治的宫颈癌组织和癌旁组织作为研究对象,采用荧光定量聚合酶链反应(PCR)分析miR-499-5p表达水平。采用对照miRNA和miR-499-5p抑制剂转染宫颈癌细胞Hela,命名为对照miRNA组和miR-499-5p KD组,采用细胞计数试剂盒(CCK-8)、克隆形成实验和5-乙炔基-2’脱氧尿嘧啶核苷(5-Ethynyl-2′-deoxyuridine,EdU)染色法分析肿瘤细胞增殖能力;采用划痕和Transwell分析两组细胞的迁移能力。采用生物信息学和双荧光素酶报告基因分析miR-499-5p靶基因。采用蛋白质免疫印迹(Western blot)分析两组细胞增殖、侵袭和靶蛋白表达水平。组间比较采用t检验。结果宫颈癌组织中miR-499-5p表达水平(1.78±0.21)明显高于癌旁组织(1.07±0.14),差异有统计学意义(t=22.920,P<0.05)。对照组细胞吸光度(A)值水平(2.23±0.15)明显高于miR-499-5p KD组(1.45±0.08),差异有统计学意义(t=11.150,P<0.05)。对照组细胞克隆形成率[(80.97±3.36)%]明显高于miR-499-5p KD组[(57.99±9.87)%],差异有统计学意义(t=5.396,P<0.05)。对照组细胞克隆形成率[(87.24±3.87)%]明显高于miR-499-5p KD组[(67.38±2.51)%],差异有统计学意义(t=22.920,P<0.05)。对照组细胞细胞核增殖抗原(Ki-67)表达水平(1.36±0.10)明显高于miR-499-5p KD组(0.92±0.06),差异有统计学意义(t=9.218,P<0.05)。对照组细胞划痕愈合率[(73.37±4.01)%]明显高于miR-499-5p KD组[(57.21±2.29)%],差异有统计学意义(t=7.637,P<0.05)。对照组细胞克隆形成率[(106.83±8.95)个]明显高于miR-499-5p KD组[(81.17±7.49)个],差异有统计学意义(t=5.384,P<0.05)。对照组细胞黏着斑激酶(FAK)蛋白表达水平(1.16±0.11)明显高于miR-499-5p KD组(0.67±0.06),差异有统计学意义(t=9.409,P<0.05)。CYLD是miR-499-5p靶基因。对照组细胞CYLD蛋白表达水平(0.98±0.08)明显低于miR-499-5p KD组(1.59±0.06),差异�Objective To investigate the effect of microRNA(miRNA,miR)-499-5p targeting CYLD on tumor cell proliferation and invasion.Methods Cervical cancer tissues and adjacent tissues admitted to our hospital from May 2020 to May 2022 were selected as the research subjects,and the expression levels of miR-499-5p were analyzed using fluorescence quantitative polymerase chain reaction(PCR).Cervical cancer cell Hela was transfected with control miRNA and miR-499-5p inhibitors,named the control miRNA group and miR-499-5p KD group.The tumor cell proliferation ability was analyzed using cell counting kit-8(CCK-8),clone formation assay,and 5-Ethynyl-2′-deoxyuridine(EdU)staining methods.The migration ability of the two groups of cells was analyzed using wounding healing and Transwell methods.The miR-499-5p target gene was analyzed by bioinformatics and double luciferase Reporter gene.The levels of cell proliferation,invasion,and target protein expression in both groups were analyzed by Western blotting.The comparison of measurement data between groups was conducted using t-test.Results The expression level of miR-499-5p in cervical cancer tissue(1.78±0.21)was significantly higher than that in adjacent tissues(1.07±0.14)(t=22.920,P<0.05).The level of cell absorbance(A)in the control group(2.23±0.15)was significantly higher than that in the miR-499-5p KD group(1.45±0.08)(t=1.150,P<0.05).The cell clone formation rate in the control group[(80.97±3.36)%]was significantly higher than that in the miR-499-5p KD group[(57.99±9.87)%,t=5.396,P<0.05].The cell clone formation rate in the control group[(87.24±3.87)%]was significantly higher than that in the miR-499-5p KD group[(67.38±2.51)%,t=22.920,P<0.05].The expression level of proliferation cell nuclear antigen(Ki-67)in the control group cells(1.36±0.10)was significantly higher than that in the miR-499-5p KD group cells(0.92±0.06,t=9.218,P<0.05).The scratch healing rate of the control group cells[(73.37±4.01)%]was significantly higher than that of the miR-499-5p KD group cells

关 键 词：微小RNA-499-5p 肿瘤进展 CYLD 增殖 侵袭 

分 类 号：R737.33[医药卫生—肿瘤]